Nelz Sophie-Charlotte, Lück Elisabeth, Schölzel Anne, Sauer Martin, Heskamp Jacqueline, Doss Sandra
Division of Nephrology, Center for Internal Medicine, Rostock University Medical Center, 18057 Rostock, Germany.
Department of Extracorporeal Therapy Systems, Fraunhofer Institute for Cell Therapy and Immunology IZI, 18057 Rostock, Germany.
Int J Mol Sci. 2025 Apr 3;26(7):3334. doi: 10.3390/ijms26073334.
hiPSC-derived blood-brain barrier (BBB) models are valuable for pharmacological and physiological studies, yet their translational potential is limited due to insufficient cell phenotypes and the neglection of the complex environment of the BBB. This study evaluates the plasma compatibility with hiPSC-derived microvascular endothelial-like cells to enhance the translational potential of in vitro BBB models. Therefore, plasma samples (sodium/lithium heparin, citrate, EDTA) and serum from healthy donors were tested on hiPSC-derived microvascular endothelial-like cells at concentrations of 100%, 75%, and 50%. After 24 h, cell viability parameters were assessed. The impact of heparin-anticoagulated plasmas was further evaluated regarding barrier function and endothelial phenotype of differentiated endothelial-like cells. Finally, sodium-heparin plasma was tested in an isogenic triple-culture BBB model with continuous TEER measurements for 72 h. Only the application of heparin-anticoagulated plasmas did not significantly alter viability parameters compared to medium. Furthermore, heparin plasmas improved barrier function without increasing cell density and induced a von Willebrand factor signal. Finally, continuous TEER measurements of the triple-culture model confirmed the positive impact of sodium-heparin plasma on barrier function. Consequently, heparin-anticoagulated plasmas were proven to be compatible with hiPSC-derived microvascular endothelial-like cells. Thereby, the translational potential of BBB models can be substantially improved in the future.
人诱导多能干细胞（hiPSC）来源的血脑屏障（BBB）模型对药理学和生理学研究具有重要价值，然而，由于细胞表型不足以及对血脑屏障复杂环境的忽视，其转化潜力受到限制。本研究评估了血浆与hiPSC来源的微血管内皮样细胞的相容性，以提高体外血脑屏障模型的转化潜力。因此，对来自健康供体的血浆样本（钠/锂肝素、柠檬酸盐、乙二胺四乙酸）和血清在hiPSC来源的微血管内皮样细胞上进行了100%、75%和50%浓度的测试。24小时后，评估细胞活力参数。进一步评估了肝素抗凝血浆对分化的内皮样细胞的屏障功能和内皮表型的影响。最后，在同基因三培养血脑屏障模型中测试了钠肝素血浆，并连续72小时测量跨内皮电阻（TEER）。与培养基相比，只有应用肝素抗凝血浆不会显著改变活力参数。此外，肝素血浆改善了屏障功能，而不增加细胞密度，并诱导了血管性血友病因子信号。最后，三培养模型的连续TEER测量证实了钠肝素血浆对屏障功能的积极影响。因此，肝素抗凝血浆被证明与hiPSC来源的微血管内皮样细胞相容。从而，未来血脑屏障模型的转化潜力可以得到显著提高。
